The effects of Bone marrow mesenchymal stem cells on m6A RNA methylation modification profile of aged granulosa cells

Abstract Background Ovarian ageing causing endocrine disturbances and degenerates systemic tissue and organ functions, seriously affects women's physical and mental health, effective treatment methods are urgently needed. On the basis of our previous studies on juvenile rhesus monkey bone marrow mesenchymal stem cells(BMMSCs) treated ovarian ageing of rhesus monkey, found that BMMSCs improved the ovarian structure and function. This study continues to explore the mechanism of BMMSCs reversd granulosa cell(GC) ageing. Methods Established GC ageing model and coculture system of BMMSCs, detected m6A methylation modification changes, performed m6A modified RNA immunoprecipitation sequencing (MeRIP-seq), corrected between m6A peaks with mRNA expression, and identified the expression of hub genes through q-PCR, Immunofluorescence staining, and western blot . Results Our results showed that H2O2 induced GC aging successfully and that BMMSCs reversed measures of GC ageing, BMMSCs promoted the expression of FTO and reduced the overall methylation level of m6A. We identified 797 m6A peaks (348 hypomethylated, 449 hypermethylated) and 817 differentially expressed genes (DEGs) (412 upregulated, 405 downregulated) after aged GCs cocultured with BMMSCs, which associated with ovarian function and epigenetic modification. Lysine Demethylase 8 (KDM8) involve in the negative regulation of osteoblast differentiation which was contracted with the positive regulation of osteoblast differentiation to mediate cellular senescence, and downregulated after treated with BMMSCs. Conclusions KDM8 may become new target in BMMSCs reversed ovarian ageing.